Poster abstracts
Poster number 28 submitted by Caroline Zu
Defining the structural basis of RNA substrate recognition by human Pus enzymes
Caroline X. Zu (Chemistry, University of Michigan), Amelia Cochran (Chemistry, University of Michigan), Markos Koutmos (Chemistry, University of Michigan)
Abstract:
Chemical modification of RNA is essential for proper cellular function, and dysregulation of RNA-modifying pathways is associated with numerous diseases, including cancer. Pseudouridine is one of the most abundant post-transcriptional RNA modifications and is installed by pseudouridine synthases (Pus enzymes) through the isomerization of uridine. Despite the widespread importance of pseudouridylation, the molecular principles governing RNA substrate recognition by Pus enzymes remain poorly understood. In particular, no structures of a eukaryotic Pus enzyme bound to its RNA substrate have been reported, limiting our understanding of how these enzymes recognize diverse RNA sequences and structures. This research employs both biochemical and structural studies to characterize the substrate scope and specificity across human Pus enzymes. The human TruB homologs TRUB1 and TRUB2 were successfully cloned into their respective expression vectors. Human PUS7 binding was quantified for RNA substrates that are either modified or not modified by PUS7. Together, this work aims to define the molecular features governing RNA substrate recognition by human Pus enzymes and establish a framework for understanding how RNA structure and sequence impact pseudouridylation specificity.
Keywords: pseudouridine, RNA modifications, Pus enzymes